{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:47:52Z","timestamp":1787028472333,"version":"build-2736575974"},"reference-count":42,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2022,3,25]],"date-time":"2022-03-25T00:00:00Z","timestamp":1648166400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005687","name":"Goethe University Frankfurt","doi-asserted-by":"publisher","award":["Not available"],"award-info":[{"award-number":["Not available"]}],"id":[{"id":"10.13039\/501100005687","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Frankfurter Stiftung f\u00fcr krebskranke Kinder","award":["Not available"],"award-info":[{"award-number":["Not available"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Mutations in the Von Hippel\u2013Lindau (VHL) gene are the driving force in many forms of clear cell renal cell carcinoma (ccRCC) and promote hypoxia-inducible factor (HIF)-dependent tumor proliferation, metastasis and angiogenesis. Despite the progress that has already been made, ccRCC generally remain resistant to conventional therapies and ccRCC patients suffer from metastasis and acquired resistance, highlighting the need for novel therapeutic options. Cysteinyl leukotriene receptor 1 (CysLTR1) antagonists, like zafirlukast, are administered in bronchial asthma to control eicosanoid signaling. Intriguingly, long-term use of zafirlukast decreases cancer risk and leukotriene receptor antagonists inhibit tumor growth, but the mechanisms still remain unexplored. Therefore, we aim to understand the mechanisms of zafirlukast-mediated cell death in ccRCC cells. We show that zafirlukast induces VHL-dependent and TNF\u03b1-independent non-apoptotic and non-necroptotic cell death in ccRCC cells. Cell death triggered by zafirlukast could be rescued with antioxidants and the PARP-1 inhibitor Olaparib, and additionally relies on HIF-2\u03b1. Finally, MG-132-mediated proteasome inhibition sensitized VHL wild-type cells to zafirlukast-induced cell death and inhibition of HIF-2\u03b1 rescued zafirlukast- and MG-132-triggered cell death. Together, these results highlight the importance of VHL, HIF and proteasomal degradation in zafirlukast-induced oxidative cell death with potentially novel therapeutic implications for ccRCC.<\/jats:p>","DOI":"10.3390\/ijms23073567","type":"journal-article","created":{"date-parts":[[2022,3,25]],"date-time":"2022-03-25T15:31:21Z","timestamp":1648222281000},"page":"3567","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Zafirlukast Induces VHL- and HIF-2\u03b1-Dependent Oxidative Cell Death in 786-O Clear Cell Renal Carcinoma Cells"],"prefix":"10.3390","volume":"23","author":[{"given":"Christopher","family":"Wolf","sequence":"first","affiliation":[{"name":"Institute for Experimental Cancer Research in Pediatrics, Goethe University Frankfurt, 60528 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sonja","family":"Smith","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cancer Research in Pediatrics, Goethe University Frankfurt, 60528 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sjoerd J. L.","family":"van Wijk","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cancer Research in Pediatrics, Goethe University Frankfurt, 60528 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"17009","DOI":"10.1038\/nrdp.2017.9","article-title":"Renal cell carcinoma","volume":"3","author":"Hsieh","year":"2017","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1038\/bjc.2016.230","article-title":"Medical treatment of renal cancer: New horizons","volume":"115","author":"Greef","year":"2016","journal-title":"Br. J. Cancer"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"15698","DOI":"10.1038\/s41598-017-15922-4","article-title":"Incidence and mortality of kidney cancer: Temporal patterns and global trends in 39 countries","volume":"7","author":"Wong","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2649","DOI":"10.1002\/cam4.767","article-title":"Projections of cancer incidence and cancer-related deaths in Germany by 2020 and 2030","volume":"5","author":"Quante","year":"2016","journal-title":"Cancer Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1038\/onc.2017.338","article-title":"The HIF and other quandaries in VHL disease","volume":"37","author":"Tarade","year":"2018","journal-title":"Oncogene"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Cancer Genome Atlas Research Network (2013). Comprehensive molecular characterization of clear cell renal cell carcinoma. Nature, 499, 43\u201349.","DOI":"10.1038\/nature12222"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1038\/ng.2891","article-title":"Genomic architecture and evolution of clear cell renal cell carcinomas defined by multiregion sequencing","volume":"46","author":"Gerlinger","year":"2014","journal-title":"Nat. Genet."},{"key":"ref_8","first-page":"107","article-title":"Negative and positive regulation of HIF-1: A complex network","volume":"1755","author":"Bardos","year":"2005","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/j.biocel.2004.08.012","article-title":"Hypoxia-inducible factor 1: Regulation by hypoxic and non-hypoxic activators","volume":"37","author":"Dery","year":"2005","journal-title":"Int. J. Biochem. Cell. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"pe25","DOI":"10.1126\/stke.3372006pe25","article-title":"HIFing the brakes: Therapeutic opportunities for treatment of human malignancies","volume":"2006","author":"Garcia","year":"2006","journal-title":"Sci. STKE"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1158\/0008-5472.CAN-12-2362","article-title":"Hyperactivated JNK is a therapeutic target in pVHL-deficient renal cell carcinoma","volume":"73","author":"An","year":"2013","journal-title":"Cancer Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1042\/BJ20080476","article-title":"Regulation of hypoxia-inducible factor-1alpha by NF-kappaB","volume":"412","author":"Kenneth","year":"2008","journal-title":"Biochem. J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1161\/01.ATV.0000258979.92828.bc","article-title":"Reactive oxygen species activate the HIF-1alpha promoter via a functional NFkappaB site","volume":"27","author":"Bonello","year":"2007","journal-title":"Arterioscler. Thromb Vasc. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"7546","DOI":"10.1128\/MCB.25.17.7546-7556.2005","article-title":"Mechanism of von Hippel-Lindau protein-mediated suppression of nuclear factor kappa B activity","volume":"25","author":"An","year":"2005","journal-title":"Mol. Cell. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1038\/21658","article-title":"Characterization of the human cysteinyl leukotriene CysLT1 receptor","volume":"399","author":"Lynch","year":"1999","journal-title":"Nature"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2691","DOI":"10.1172\/JCI97946","article-title":"Leukotriene receptors as potential therapeutic targets","volume":"128","author":"Yokomizo","year":"2018","journal-title":"J. Clin. Investig."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1007\/s11882-015-0564-7","article-title":"Antileukotrienes in upper airway inflammatory diseases","volume":"15","author":"Cingi","year":"2015","journal-title":"Curr. Allergy Asthma Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1016\/0140-6736(91)90277-V","article-title":"Effect of cysteinyl-leukotriene receptor antagonist ICI 204.219 on allergen-induced bronchoconstriction and airway hyperreactivity in atopic subjects","volume":"337","author":"Taylor","year":"1991","journal-title":"Lancet"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1038\/nrc2809","article-title":"Eicosanoids and cancer","volume":"10","author":"Wang","year":"2010","journal-title":"Nat. Rev. Cancer"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"23979","DOI":"10.1038\/srep23979","article-title":"Cysteinyl Leukotriene Receptor Antagonists Decrease Cancer Risk in Asthma Patients","volume":"6","author":"Tsai","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_21","first-page":"185","article-title":"The cysteinylLT1 receptor in human renal cell carcinoma","volume":"1","author":"Kawahito","year":"2008","journal-title":"Mol. Med. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1053\/gast.2003.50011","article-title":"Expression of the leukotriene D4 receptor CysLT1, COX-2, and other cell survival factors in colorectal adenocarcinomas","volume":"124","author":"Ohd","year":"2003","journal-title":"Gastroenterology"},{"key":"ref_23","first-page":"99","article-title":"Overexpression of cysteinyl LT1 receptor in prostate cancer and CysLT1R antagonist inhibits prostate cancer cell growth through apoptosis","volume":"18","author":"Matsuyama","year":"2007","journal-title":"Oncol. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"798","DOI":"10.1139\/cjpp-2017-0757","article-title":"Cysteinyl leukotriene receptor antagonists induce apoptosis and inhibit proliferation of human glioblastoma cells by downregulating B-cell lymphoma 2 and inducing cell cycle arrest","volume":"96","author":"Piromkraipak","year":"2018","journal-title":"Can. J. Physiol. Pharmacol."},{"key":"ref_25","first-page":"4199","article-title":"Chemoprevention by lipoxygenase and leukotriene pathway inhibitors of vinyl carbamate-induced lung tumors in mice","volume":"62","author":"Gunning","year":"2002","journal-title":"Cancer Res."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Savari, S., Liu, M., Zhang, Y., Sime, W., and Sj\u00f6lander, A. (2013). CysLT(1)R antagonists inhibit tumor growth in a xenograft model of colon cancer. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0073466"},{"key":"ref_27","first-page":"833","article-title":"Leukotriene Receptor Antagonists Inhibit Mitogenic Activity in Triple Negative Breast Cancer Cells","volume":"19","author":"Suknuntha","year":"2018","journal-title":"Asian Pac. J. Cancer Prev. APJCP"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"950","DOI":"10.1177\/2472555217706478","article-title":"An Innovative High-Throughput Screening Approach for Discovery of Small Molecules That Inhibit TNF Receptors","volume":"22","author":"Lo","year":"2017","journal-title":"SLAS Discov."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1002\/JLB.2AB0420-572RR","article-title":"Quantitative single-molecule imaging of TNFR1 reveals zafirlukast as antagonist of TNFR1 clustering and TNFalpha-induced NF-kB signaling","volume":"109","author":"Weinelt","year":"2021","journal-title":"J. Leukoc. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e31","DOI":"10.3346\/jkms.2020.35.e31","article-title":"Involvement of the TNF-\u03b1 Pathway in TKI Resistance and Suggestion of TNFR1 as a Predictive Biomarker for TKI Responsiveness in Clear Cell Renal Cell Carcinoma","volume":"35","author":"Hwang","year":"2020","journal-title":"J. Korean Med. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1146\/annurev-biochem-060310-170328","article-title":"The ubiquitin code","volume":"81","author":"Komander","year":"2012","journal-title":"Annu. Rev. Biochem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.molmed.2017.11.002","article-title":"Checkpoints in TNF-Induced Cell Death: Implications in Inflammation and Cancer","volume":"24","author":"Annibaldi","year":"2018","journal-title":"Trends Mol. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1038\/s41573-020-0076-6","article-title":"Poly(ADP-ribose) polymerase inhibition: Past, present and future","volume":"19","author":"Curtin","year":"2020","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/j.ctrv.2018.07.009","article-title":"Current and emerging therapies for first-line treatment of metastatic clear cell renal cell carcinoma","volume":"70","author":"Atkins","year":"2018","journal-title":"Cancer Treat. Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3322\/caac.21590","article-title":"Cancer statistics, 2020","volume":"70","author":"Siegel","year":"2020","journal-title":"CA. Cancer J. Clin."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1016\/j.biopha.2018.12.064","article-title":"Cysteinyl leukotriene receptor type 1 (CysLT1R) antagonist zafirlukast protects against TNF-alpha-induced endothelial inflammation","volume":"111","author":"Zhou","year":"2019","journal-title":"Biomed. Pharmacother."},{"key":"ref_37","first-page":"765","article-title":"Reactive oxygen species induced by icaritin promote DNA strand breaks and apoptosis in human cervical cancer cells","volume":"41","author":"Chen","year":"2019","journal-title":"Oncol. Rep."},{"key":"ref_38","first-page":"1195","article-title":"Oxidative DNA damage: Mechanisms, mutation, and disease","volume":"17","author":"Cooke","year":"2003","journal-title":"FASEB J. Off. Publ. Fed. Am. Soc. Exp. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1038\/nrc3399","article-title":"DNA repair dysregulation from cancer driver to therapeutic target","volume":"12","author":"Curtin","year":"2012","journal-title":"Nat. Rev. Cancer"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1433","DOI":"10.1093\/carcin\/bgs132","article-title":"PARPs and the DNA damage response","volume":"33","author":"Sousa","year":"2012","journal-title":"Carcinogenesis"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1615\/CritRevEukaryotGeneExpr.2013006875","article-title":"Review of poly (ADP-ribose) polymerase (PARP) mechanisms of action and rationale for targeting in cancer and other diseases","volume":"24","author":"Morales","year":"2014","journal-title":"Crit. Rev. Eukaryot. Gene Expr."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/7\/3567\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:43:02Z","timestamp":1760136182000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/7\/3567"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,25]]},"references-count":42,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,4]]}},"alternative-id":["ijms23073567"],"URL":"https:\/\/doi.org\/10.3390\/ijms23073567","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,25]]}}}